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Background Human induced pluripotent stem cells (hiPSCs) and their differentiated cell types have a great potential for tissue repair and regeneration. While the primary focus of using hiPSCs has historically been to regenerate damaged tissue, emerging studies have shown a more potent effect of hiPSC-derived paracrine factors on tissue regeneration. However, the precise contents of the transplanted hiPSC-derived cell secretome are ambiguous. This is mainly due to the lack of tools to distinguish cell-specific secretome from host-derived proteins in a complex tissue microenvironment in vivo. Methods In this study, we present the generation and characterization of a novel hiPSC line, L274G-hiPSC, expressing the murine mutant methionyl-tRNA synthetase, L274G Mm MetRS, which can be used for tracking the cell specific proteome via biorthogonal non-canonical amino acid tagging (BONCAT). We assessed the trilineage differentiation potential of the L274G-hiPSCs in vitro and in vivo. Furthermore, we assessed the cell-specific proteome labelling in the L274G-hiPSC derived cardiomyocytes (L274G-hiPSC-CMs) in vitro following co-culture with wild type human umbilical vein derived endothelial cells and in vivo post transplantation in murine hearts. Results We demonstrated that the L274G-hiPSCs exhibit typical hiPSC characteristics and that we can efficiently track the cell-specific proteome in their differentiated progenies belonging to the three germ lineages, including L274G-hiPSC-CMs. Finally, we demonstrated cell-specific BONCAT in transplanted L274G-hiPSC-CMs. Conclusion The novel L274G-hiPSC line can be used to study the cell-specific proteome of hiPSCs in vitro and in vivo, to delineate mechanisms underlying hiPSC-based cell therapies for a variety of regenerative medicine applications.

Fluorides are emitted in both gaseous and particle forms in the industrial sector. However, studies usually only report total fluoride content. Therefore, the study aimed to assess the particulate, gaseous fluoride and correlate it with the respirable dust particles in Single Super Phosphate (SSP), Granular Single Super Phosphate (GSSP), and administration divisions of the industry. Respirable dust particles, particulate fluoride, and hydrogen fluoride in the work environment were collected on a filter cassette containing an MCE filter paper (0.8 micron 37-mm) and Na2CO3 impregnated backup pad, respectively, using a personal sampler. The fluoride samples were analyzed using Ion Selective Electrode (ISE) and expressed as milligrams per meter cube (mg.m-3). The respirable dust, particulate, and gaseous fluoride content were found to have statistically significant differences (p<0.001) between the divisions (SSP, GSSP, and administration) in the static monitoring, whereas, in the case of personal monitoring, no significant differences were observed. Average airborne respirable, particulate, and gaseous fluoride levels in static monitoring were 1.37, 1.03, 0.20 mg.m-3, 0.018, 0.008, 0.001 mg.m-3, and 0.808, 0.403, 0.026 ppm in SSP, GSSP and administration respectively, whereas in personal monitoring the average respirable, particulate and gaseous fluoride concentrations were 1.18, 0.85, 0.30 mg.m-3, 0.0013, 0.007, 0.002 mg.m-3 and 0.356, 0.258, 0.011 ppm in SSP, GSSP and administration respectively. The present study observed that the levels of fluoride decreased with an increase in distance from SSP, followed by GSSP and administration. It indicates that the fluoride exposure was inversely proportional to the distance of the source. This study outcome will help to design a policy and intervention to mitigate fluoride exposure among workers.

The European Prospective Investigation into Cancer and Nutrition-Oxford(1), Adventist Health Study-2(2), and UK Biobank(3) have reported that vegetarians (including vegans) had a lower risk for total cancer incidence compared to meat-eaters. [...]the numbers of vegetarians in these studies were too low to reliably estimate the associations with site-specific cancers. [...]we established the Health in Vegetarians Consortium and here we compare the baseline dietary and anthropometric characteristics between diet groups and between cohorts. Participants were categorised into eight diet groups based on dietary questionnaires completed at recruitment. 2.3 million participants were included; 66.2% women and 33.8% men, with average ages at recruitment being 56.9 (SD: 7.8) and 57.3 (8.6) years, respectively. 1.07 million participants were regular meat-eaters, 1.06 million low meat-eaters, 60,679 poultry eaters, 43,933 pescatarians, 48,162 lacto-ovo, 18,093 lacto, 13,702 ovo vegetarians, and 13,885 vegans.

Background: IgE-reactive proteins have been identified in almond; however, few have been cloned and tested for specific patient IgE reactivity. Here, we clone and express prunin 1 and prunin 2, isoforms of the major almond protein prunin, an 11S globulin, and assay each for IgE reactivity. Methods: Prunin isoforms were PCR-amplified from an almond cDNA library, sequenced, cloned and expressed in Escherichia coli. Reactivity to the recombinant (r) allergens, Pru du 6.01 and Pru du 6.02, was screened by dot blot and immunoblot assays using sera from almond-allergic patients and murine monoclonal antibodies (mAbs). Sequential IgE-binding epitopes were identified by solid-phase overlapping peptide analysis. Epitope stability was assessed by assaying denatured recombinant proteins by immunoblot. Results: IgE reactivity to rPru du 6.01 and rPru du 6.02 was found in 9 of 18 (50%) and 5 of 18 patients (28%), respectively. Four patients (22%) demonstrated reactivity to both isoforms. Murine anti-almond IgG mAbs also showed greater reactivity to rPru du 6.01 than to rPru du 6.02. Both stable and labile epitopes were detected. Six IgE-binding sequential epitope-bearing peptide segments on Pru du 6.01 and 8 on Pru du 6.02 were detected using pooled almond-allergic sera. Conclusions: rPru du 6.01 is more widely recognized than rPru du 6.02 in our patient population. The identification of multiple sequential epitopes and the observation that treatment with denaturing agents had little effect on IgE-binding intensity in some patients suggests an important role for sequential epitopes on prunins.

Allergic reactions to tree nuts can be serious and life threatening. Considerable research has been conducted in recent years in an attempt to characterize those allergens that are most responsible for allergy sensitization and triggering. Both native and recombinant nut allergens have been identified and characterized and, for some, the IgE-reactive epitopes described. Some allergens, such as lipid transfer proteins, profilins, and members of the Bet v 1-related family, represent minor constituents in tree nuts. These allergens are frequently cross-reactive with other food and pollen homologues, and are considered panallergens. Others, such as legumins, vicilins, and 2S albumins, represent major seed storage protein constituents of the nuts. The allergenic tree nuts discussed in this review include those most commonly responsible for allergic reactions such as hazelnut, walnut, cashew, and almond as well as those less frequently associated with allergies including pecan, chestnut, Brazil nut, pine nut, macadamia nut, pistachio, coconut, Nangai nut, and acorn.

The potential epitopes of a recombinant food allergen protein, cashew Ana o 2, reactive to polyclonal antibodies, were mapped by solution-phase amide backbone H/D exchange (HDX) coupled with Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS). Ana o 2 polyclonal antibodies were purified in the serum from a goat immunized with cashew nut extract. Antibodies were incubated with recombinant Ana o 2 (rAna o 2) to form antigen:polyclonal antibody (Ag:pAb) complexes. Complexed and uncomplexed (free) rAna o 2 were then subjected to HDX-MS analysis. Four regions protected from H/D exchange upon pAb binding are identified as potential epitopes and mapped onto a homologous model. Figure ᅟ

Background: We recently cloned and described a vicilin and showed it to be a major cashew allergen. Additional IgE-reactive cashew peptides of the legumin group and 2S albumin families have also been reported. Here, we attempt to clone, express and characterize a second major cashew allergen. Methods: A cashew cDNA library was screened with human IgE and rabbit IgG anti-cashew extract antisera, and a reactive nonvicilin clone was sequenced and expressed as a fusion protein in Escherichia coli. Immunoblotting was used to screen for reactivity with patients’ sera, and inhibition of immunoblotting was used to identify the corresponding native peptides in cashew nut extract. The identified allergen was subjected to linear epitope mapping using SPOTs solid-phase synthetic peptide technology. Results: Sequence analysis showed the selected clone, designated Ana o 2, to encode for a member of the legumin family (an 11S globulin) of seed storage proteins. By IgE immunoblotting, 13 of 21 sera (62%) from cashew-allergic patients were reactive. Immunoblot inhibition data showed that the native Ana o 2 constitutes a major band at approximately 33 kD and a minor band at approximately 53 kD. Probing of overlapping synthetic peptides with pooled human cashew-allergic sera identified 22 reactive peptides, 7 of which gave strong signals. Several Ana o 2 epitopes were shown to overlap those of the peanut legumin group allergen, Ara h 3, in position but with little sequence similarity. Greater positional overlap and identity was observed between Ana o 2 and soybean glycinin epitopes. Conclusions: We conclude that this legumin-like protein is a major allergen in cashew nut.

Spinal anaesthesia requires a small volume of drug to produce profound reproducible sensory analgesia and motor blockade, but has limited duration of action. A properly chosen adjuvant to local anaesthetic agent produces the best way to achieve a better quality regional block. Hence, a study was conducted to compare the effect of intrathecal clonidine 75 μg or neostigmine 50 μg added to intrathecal hyperbaric bupivacaine, with regards to sensory characteristics, motor characteristics, haemodynamic stability and side effects. This was a prospective randomized experimental study in 50 patients posted for lower abdominal surgery belonging to ASA I and II status and aged between18 and 60 years. One group received intrathecal clonidine 75 μg and 2.5 ml (12.5 mg) of intrathecal 0.5% hyperbaric bupivacaine (group BC) and second group received neostigmine 50 μg with 2.5 ml (12.5mg) of intrathecal 0.5% hyperbaric bupivacaine (group BN) and they were compared with regards to sensory characteristics, motor characteristics, haemodynamic stability and side effects. Addition of 50 μg neostigmine significantly enhanced the onset of sensory block (BN - 90 ± 15 secs, BC-160 ± 20 secs, P value as <0.05) and motor block (BN-110 ± 15 secs, BC-210 ± 20 secs, P value as <0.05) compared to clonidine. Haemodynamics were well maintained in the neostigmine group. Group BC had prolonged analgesia (362 ± 36 mins) compared to BN group (300 ± 25 mins)(P < 0.05) with no serious adverse effects noted perioperatively in either groups. Intrathecal clonidine with hyperbaric bupivacaine produces prolonged postoperative analgesia and intrathecal neostigmine with bupivacaine produces a good sensory and motor for the surgical procedure.